Human embryonic stem cells and microenvironment

Abstract

Human embryonic stem cells (hESCs) possess a great potential in the field of regenerative medicine by their virtue of pluripotent potential with indefinite proliferation capabilities. They can self renew themselves and differentiate into three embryonic germ layers. Although they are conventionally grown on mitotically inactivated mouse feeder cells, there are in vitro culture systems utilizing feeder cells of human origin in order to prevent cross-species contamination. Recently established in vitro culture systems suggested that direct interaction with feeder cells is not necessary but rather attachment to a substrate is required to ensure long-term, efficient hESC culture in vitro. This substrate is usually composed of a mixture of extracellular matrix components representing in vivo natural niche. In hESC biology, the mechanism of interaction of hESCs with extracellular matrix molecules remained insufficiently explored area of research due to their transient nature of interaction with the in vivo niche. However, an in vitro culture system established using extracellular matrix molecules may provide a safer alternative to culture systems with feeder cells while paving the way to Good Manufacturing Practice-GMP production of hESCs for therapeutic purposes. Therefore, it is essential to study the interaction of extracellular matrix molecules with hESCs in order to standardize in vitro culture systems for large-scale production of hESCs in a less labor-intensive way. This would not only provide valuable information regarding the mechanisms that control pluripotency but also serve to dissect the molecular signaling pathways of directed differentiation for prospective therapeutic applications in the future. J Clin Exp Invest 2014; 5 (3): 486-495

Keywords

• Human embryonic stem cells, extracellular matrix, pluripotency, in vitro culture systems.

İnsan embriyonik kök hücreleri ve mikroçevre

Öz

İnsan embriyonik kök hücreleri (iEKH) pluripotent özellikleri ile sınırsız çoğalıp kendi kendilerini yenileyebilirken, üç embriyonik tabakayı temsil eden hücrelere farklılaşabilme yetenekleri ile rejeneratif tıp alanında büyük ilgi uyandırmışlardır. Genellikle mitotik olarak inaktive edilmiş fare besleyici hücreleri ile büyütülseler de, türler arası kontaminasyon riskini ortadan kaldırmak için insan kökenli besleyici hücrelerin kullanıldığı in vitro kültür sistemleri de bulunmaktadır. Son dönemde geliştirilen kültürlerde iEKH\'nin besleyici hücre ile birebir temasına gerek olmadığı, ancak hücre tutunmasını sağlayan substratın varlığının uzun süreli, etkin in vitro iEKH\'nin kültürü için gerekli olduğu ileri sürülmüştür. Bu substrat çoğunlukla in vivo mikroçevrenin de bir parçası olan ekstrasellüler matriks moleküllerinden biri ya da birkaçının karışımı olabilmektedir. İEKH biyolojisinde ekstrasellüler matriks moleküleriyle etkileşim, hücrelerin kısa süreli ve geçici in vivo mikroçevre ile interaksiyonu nedeniyle şimdiye kadar ihmal edilen bir alan olarak kalmıştır. Ancak ekstraselüler matriks molekülleriyle oluşturulacak bir in vitro kültür sistemi, besleyici hücrelerin kullanıldığı geleneksel kültür sistemlerine nazaran güvenli bir alternatif oluşturarak, ‘İyi Üretim Uygulamaları (GMP)\' standardında tedaviye yönelik iEKH\'nin üretiminin yolunu açacaktır. Bu nedenle iEKH\'nin geniş çaplı üretiminin, iş yükünü en aza indirecek şekilde yapılabilmesi için gerekli in vitro kültür standartlarının oluşturulması, iEKH\'nin ekstraselüler matriks molekülleriyle ilişkilerinin araştırılmasına bağlıdır. Böylelikle hem pluripotent özelliği kontrol eden mekanizmalar hakkında önemli bilgi edinilirken hem de gelecekteki tedaviye yönelik olası uygulamalarda kullanılacak iEKH\'nin yönlendirilmiş farklılaşmasını sağlayan sinyal yolakları tanımlanabilecektir.

Anahtar Kelimeler

• İnsan embriyonik kök hücreleri, ekstraselüler matriks, pluripotent özellik, in vitro kültür sistemleri.

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